Australian Journal of Basic and Applied Sciences

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1 Australian Journal of Basic and Applied Sciences, 8() July 14, Pages: AENSI Journals Australian Journal of Basic and Applied Sciences ISSN: Journal home page: Variations of Stem Anatomy of Cabbage Seedlings (Brassica oleracea L.) Seedlings Produced from Heat-Shock Callus Tissue Jamella H. Rasheed, Faiq H. Alradi, Sahla M. Alzeadan Biology department, Education college for Pure Sciences, Mosul University, Mosul. IRAQ. A R T I C L E I N F O Article history: Received April 14 Received in revised form 8 May 14 Accepted May 14 Available online 17 June 14 Keywords: Brassica olerea, stem anatomy heat-shock A B S T R A C T Background: The current study concentrated from an effect of heat- shock exposure of callus derived from stem segments of cabbage plants (Brassica olerea L.). Objective: The heat shock (,,0,5 and Cº) for and minutes for each degree increased callus growth, size and easy to regenerate the callus to shoot and root. Results: Regeneration ratio reach up to 91.66%, especially in temperature Cº for minutes, and the numbers of shoots were 1, while in the same degree for minutes the ratio of regeneration was 44.2% and the shoots were 2. On the other hand the anatomical characteristics are different in the shape, numbers of vascular bundles and the pith area. Conclusion: This study aimed to identify the anatomical variation in stem of seedlings produced from callus exposed to heat-shock. 14 AENSI Publisher All rights reserved. To Cite This Article: Jamella H. Rasheed, Faiq H. Alradi, Sahla M. Alzeadan., Variations of Stem Anatomy of Cabbage Seedlings (Brassica oleracea L.) Seedlings Produced from Heat-Shock Callus Tissue. Aust. J. Basic & Appl. Sci., 8(): 84-88, 14 INTRODUCTION Plant biotechnology its applications, are an important tool for breeding activity toward the enhancement of qualitative and quantitative traits of these important vegetable (Cristea et al.12). Brassicaceae contain substantial quantities of bioactive compounds, which are good free radical scavenger and might have strong antitumor properties. (Caker, et al., 12). Climatic factors, such as extreme temperature (heat, cold and freezing), are major abiotic environmental stress that limit plant growth and development. Therefor agronomical yield and had a major role in determining the geographic distribution of plant species. (Krasenky & Jonak, 12). The positive heat treatments in some systems of plants, stimulating metabolism ways in the cell presented in building up and accumulation of new proteins called (HSP) heat shock protein.(lee et al., 00). In addition the evidence due to that heat shock is inducting the oxidative operations and promote of synthesizing the enzymes antioxidant in number of plants. Such as the exposure of sunflower cell suspension culture to Cº for hours. As well as to recognize on the internal anatomical variations happens in tissue culture plants compared with the same plants produced from seeds clear differences in the anatomical features of Cucuis milo L. plants were produced from tissue culture including enlargement of diameter of stem and numbers of vascular bundles compared with seed plants. (Mavrona et al., 00), and specific anatomical variation in epidermis and cortex in Citrullus lanatus plants and Largenaria siceraria (Yetisire et al., 05). This study aimed to identify the anatomical variation in stem of seedlings produced from callus exposed to heat-shock. MATERIAL AND METHODS Sterile seedling production: Cabbage's seed (Brassica oleracea L.) were sterilized by soaking it in 6% sodium hypochloride solution (1:2 v/v) for minutes, and washed the seed thoroughly by sterile water. Sterilized seeds were transported to the surface of ml MS0 (Murashige & Skoog,19) medium contained in 0 ml( seeds/flask), specimens were kept in culture room at condition of dark at ± 2Cº, then the grown seedlings were transported to light and dark successive conditions 16/8 hours at light intensity 1500 lux. Exposure of callus to heat shock: One gram sample of stem derived callus 4 weeks old were each in sterile tube with cover of ml capacity. Samples were exposed to each at (,, 0, 5, and Cº) for and minutes. Each exposed test tube was Corresponding Author: Faiq H. Alradi, Biology department, Education college for Pure Sciences, Mosul University, Mosul. IRAQ. Ph: ( ) faiq_alradi7@yahoo.com.

2 85 Rasheed et al, 14 Australian Journal of Basic and Applied Sciences, 8() July 14, Pages: directly in water at roof temperature in waterbath, at the selected degree previously fixed (Irina et al., 02). All heat- treated samples were placed on agar, solidified MS medium supplemented with 1.0 mg/l NAA+ 2.0 mg/l BA and incubated at the same conditions(alradi, 1). Preparation of sections: The basic steps of sample fixation, embedding in wax were followed as described (Al-haj, 1998) to prepare permanent various sections. Results: Effect of heat shock on callus stimulation from explants: The results in table (1) indicate that all treatment enhanced callus stimulation. Table 1: Effect of some treatment of heat- shock on callus production from stem explants of cabbage (Brassica oleracea L.). Treatments Cº/min. No. of stem cultured Explants responded Callus % In addition the images in figure.1 showed of the beginning callus stimulated is derived from stem egments of cabbage plants (Brassica oleracea L.) in solid MS media is provided with 1.0 mg/l NAA mg/l BA (figure1-a). In the other hand stimulated callus which exposure to heat shock in optimum temperature at Cº for minutes to show increasing size of callus (figure 1-B), and the shoot is in initiating from callus (figure 1- C). Fig. 1: A-Callus initiation from cabbage's stem, B-Increasing callus in size after exposure to heat-shock, C- Start shoots from callus. Table(2) show the effects of treatments heat-shock on callus regeneration and the number of shoots, it was the maximum at Cº for minutes to arrival the segments regenerated 22, the number of shoots 1, and the ratio of regeneration was Table 2: Effect of some treatments of heat-shock on regeneration of callus of cabbage Brassica oleracea. Treatments No. of callus Segment No. of Cº/min. exposed regenerated shoots Regeneration (%)

3 86 Rasheed et al, 14 Australian Journal of Basic and Applied Sciences, 8() July 14, Pages: In the figure-2 images indicate number of shoots (figure2-a), and complete regeneration to hole plant under tissue culture conditions (figure2-b). The plant transported to the soil after 0 day from callus is begging regeneration (figure2-c). Fig. 2: A-Number shoots regenerated of callus, B- Complete regeneration to shoots and roots, D- Adaptation the plant by transported to the soil. The anatomical changes in cross sections of stems: Cross section of stem derived from callus exposure to heat- shock (figure.), is bigger than cross section of stem in seed plants (figure.4), in size and its more folded or curvatures. The stomatal system is looking wide (figure.-a), compared with in (figure.4-a). In the cells cortex showed as corner cholenchyma (figure.-b), but is absence in another (figure.-b). The numbers of vascular bundles in cross section of stem of tissue culture 9 bundle/section. (figure.-c), while being 7 bundle/section in cross section of stem in seed plants. (figure.4-c), as well as the pith zone is wide (figure.-d), compared with another (figure.4-d). Fig. : C.S. cabbage's stem derived from callus exposure to heat- sho. Fig. 4: C.S. cabbage's stem derived from seed plants.

4 87 Rasheed et al, 14 Australian Journal of Basic and Applied Sciences, 8() July 14, Pages: The more studies had proven on role of using the heat shock in promoting some biotic activities in the cell. (Sholpan et al., 01). Callus increase in size which is exposure to heat treatments, maybe is attributable to enhancement of protein and chlorophyll in the cells of callus, because of the heat shock. (Al-Mallah & Al- Nema, 07). The positive role in obtaining to increase permeability of cellular membranes, which is to attract Ca +2 and some nutrients. (Mejia et al., 1995). A study found that exposure the cell suspension for heat shock lead to augment cells divisions in rice plants. (Thompson et al., 1989), as well as that lead to early cells divisions and overmuch of the numbers in callus permodia is initiating from cell suspension to sunflower plants. (Rasheed & Kasim, 06). The effect of promoting to this treatment is representing to remove all the barriers in the cell membrane through formatting temporary pores. (Joersbo & Brunstedt, 1991). In high temperature lead to decrease of regenerating for callus to shoot and root, that perhaps return to obtain damage in proteins of the cells. (Bukner et al., 1998), or is crashing the responsible enzymes of it building. (Soloman et al., 1999), that is consequent cells death. (Loschiavo et al., 00). This study focused on the anatomical characteristics, because of the value to recognize the changes as reason to culture the plants under in vitro conditions, environmental conditions effected on the anatomical features to enhance are desirable features to produce plant is able in resistance the worse conditions. (Edwin et al., 08). In the other hand a study found the numbers of vascular bundles increase compared with the same plant is culturing in the soil. (Alradi, 1). REFERENCES Al-haj, H.A., The light microbial preparation. Jordan. Al-Mallah, M.K. and K.S. Al-Nema, 07. Heat shock increasing protein and chlorophyll content in callus of Dianthus caryophyllus L. Edu. & Sci. J., : Alradi, F.H., 1. Anatomical-histo study on level of light & transmission electron microscope of Fenugreek plants Trigonella foenum-graecum L. regenerated from tissue culture and its hairy roots transformed by Agrobacterium rhizogenes R11. Ph.D. Mosul. Iraq. Bukner, B., D. Janick-Buckner, J. Gary and G.M. Johal, Cell death mechanisms in maize. Trends Plant Sci., : Caker, J., A. Paric, M. Maksimovic and K. Bajrovic, 12. Antioxidative and antitumor properties of in vitro cultivated broccoli (Brassica oleracea var. italia). Pharm Biol., 50: Cristea, T.O., C. Leonte, M. Prisecaru, C. Brezeanu and M. Brezeanu, 12. Effect of carbohydrate source over the androgenesis of Brassica oleracea L. anther cultivated in vitro. Seria Agronomi J., 55: Edwin, F.G., A.H. Michael and D. Geert-Jan, 08. Plant propagation by tissue culture rd edition, 1. Springer. Irina, I.P., A.V. Roman and S. Friedrich, 02. Heat stress and heat shock transcription factor-dependent expression and activity of ascorbate peroxidase in Arabidopsis. Plant Physiol., 129: Joersbo, M. and J. Brunstedt, Stimulation of protein synthesis in elecctroporated plant protoplast. J. Plant Physiol., 16: Krassensky, J. and C. Jonak, 12. Drought, salt and temperature stress- induced metabolic rearrangements and regulatory networks. J. Exp. Bot., 1-6. Lee, B.H., S.H. Won, S. Lee and M. Miyao, 00. Expression of the chloroplast- localized small heat shock protein by oxidative stress in rice. Gene. Bio., 245: Loschiavo, F., B. Balan, D. Compagnin, R. Ganz and P. Mariani, 00. Spontaneous and induced apoptosis in embryogenic cell cultures of carrot (Daucus carota) indifferent physiological state. Eu. J. Cell. Bio., 79: Mavrona, E.T., M.K. Sotiria and T. Pritsa, 00. Response of squash (Cucurbita spp.) as root stock for melon (Cucumis melo L.). J. Sci. Cult., 8: 5-. Mejia, R., E. Gomez and M.S. Fememez, Membrance fluidity of plant cell during heat schock. Biochem. Biophys. Acta., 129: Mita, G., G. Nocco, C. Leuci, V. Grco, P. Rampiro and C. Perrotta, Secreted heat shock proteins in sunflower suspension cell cultures. Plant Cell Repts., 16: Murashige, T. and F. Skoog, 19. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant, 15: Rasheed, J.H. and W.S. Kasim, 06. The role of heat treatment in inducting cellular division and callus formation from cell suspension of sunflower (Helianthus annuus) plants in multiple agar drops. Raf. Agri. J., 4: Sholpan, D., M. Tamas, M. Pal, O. Attila, T. Katalin, M. Zoltan, D. Denes and D. Maria, 01. Auxin and heat shock activation of novel member of the calmodulin like domain protein kinase gene family in cultured alfalfa cells. J. Exp. Bot., 52: Soloman, M., B. Belenghi, Delledon and E. Menachem, The involvement of cysteine proteases and protease inhibitor genes in the regulation of programmed cell death in plant. Plant Cell Repts., 11:

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